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Biochemical oxygen demand
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=== Biosensor === An alternative to measure BOD is the development of biosensors, which are devices for the detection of an analyte that combines a biological component with a physicochemical detector component. Enzymes are the most widely used biological sensing elements in the fabrication of biosensors. Their application in biosensor construction is limited by the tedious, time-consuming and costly enzyme purification methods. Microorganisms provide an ideal alternative to these bottlenecks.<ref>{{cite web |last1=Lei |first1=Yu |title=Microbial biosensors |url=http://www.cbs.umn.edu/sites/default/files/public/downloads/microbial%20sensor%2006.pdf |website=www.cbs.umn.edu |publisher=Analytica Chimica Acta 568 (2006) 200β210 |access-date=2014-09-16 |archive-url=https://web.archive.org/web/20150319212940/http://www.cbs.umn.edu/sites/default/files/public/downloads/microbial%20sensor%2006.pdf |archive-date=2015-03-19 |url-status=dead }}</ref> Many micro organisms useful for BOD assessment are relatively easy to maintain in pure cultures, grow and harvest at low cost. Moreover, the use of microbes in the field of biosensors has opened up new possibilities and advantages such as ease of handling, preparation and low cost of device. A number of pure cultures, e.g. ''Trichosporon cutaneum, Bacillus cereus, Klebsiella oxytoca, Pseudomonas sp.'' etc. individually, have been used by many workers for the construction of BOD biosensor. On the other hand, many workers have immobilized activated sludge, or a mixture of two or three bacterial species and on various membranes for the construction of BOD biosensor. The most commonly used membranes were polyvinyl alcohol, porous hydrophilic membranes etc.<ref name="igib.res.in">{{cite web |last1=Kumar |first1=Rita |title=Immobilized Microbial Consortium Useful for Rapid and Reliable BOD Estimation |url=http://www.igib.res.in/pme/patents.htm |date=2004 |website=Patents |publisher=CSIR-Institute of Genomics & Integrative Biology (IGIB) |location=New Delhi, India |id=United Kingdom; GB2360788;(3-11-2004)}}</ref> A defined microbial consortium can be formed by conducting a systematic study, i.e. pre-testing of selected micro-organisms for use as a seeding material in BOD analysis of a wide variety of industrial effluents. Such a formulated consortium can be immobilized on suitable membrane, i.e. charged nylon membrane. Charged nylon membrane is suitable for microbial immobilization, due to the specific binding between negatively charged bacterial cell and positively charged nylon membrane. So the advantages of the nylon membrane over the other membranes are : The dual binding, i.e. Adsorption as well as entrapment, thus resulting in a more stable immobilized membrane. Such specific Microbial consortium based BOD analytical devices, may find great application in monitoring of the degree of pollutant strength, in a wide variety of industrial waste water within a very short time.<ref name="igib.res.in" /> Biosensors can be used to indirectly measure BOD via a fast (usually <30 min) to be determined BOD substitute and a corresponding calibration curve method (pioneered by Karube et al., 1977). Consequently, biosensors are now commercially available, but they do have several limitations such as their high maintenance costs, limited run lengths due to the need for reactivation, and the inability to respond to changing quality characteristics as would normally occur in wastewater treatment streams; e.g. diffusion processes of the biodegradable organic matter into the membrane and different responses by different microbial species which lead to problems with the reproducibility of result (Praet et al., 1995). Another important limitation is the uncertainty associated with the calibration function for translating the BOD substitute into the real BOD (Rustum ''et al.'', 2008).
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